Lipids and compositions for intracellular delivery of biologically active compounds

Abstract

The present invention provides novel amino-lipids, compositions comprising such amino-lipids and methods of producing them. In addition, lipid nanoparticles comprising the novel amino-lipids and a biologically active compound are provided, as well as methods of production and their use for intracellular drug delivery.

What is claimed is: 
     
       1. A cyclic amino-lipid of the formula: 
       
         
           
           
               
               
           
         
         wherein: 
         R1 is independently selected from
 —(CH 2 ) 2 —N(R) 2 ; 
 —(CH 2 ) 2 —N(R)—(CH 2 ) 2 —N(R) 2 , wherein R is independently selected from —H, 
 C6-40 alkyl, C6-40 alkenyl and C6-C40 alkynyl, provided that —N(R) 2  is not NH 2 ; 
 C6-40 alkyl, and C6-40 alkenyl; and 
 
         R2 is C6-40 alkyl, C6-40 alkenyl or C6-40 alkynyl; and 
         m is 0 or 1, provided that when m is 0, R1 is not C6-C40 alkyl or C6-C40 alkenyl; 
         and pharmaceutically acceptable salts thereof. 
       
     
     
       2. The cyclic amino-lipid of  claim 1 , selected from the group of 
       
         
           
           
               
               
           
         
       
     
     
       3. A lipid nanoparticle comprising a cyclic amino-lipid of  claim 1 . 
     
     
       4. The lipid nanoparticle of  claim 3 , further comprising an additional lipid selected from the group consisting of: a cationic lipid, a helper lipid, and a PEG-lipid. 
     
     
       5. The lipid nanoparticle of  claim 3 , further comprising a hydrophobic small molecule. 
     
     
       6. The lipid nanoparticle of  claim 3 , further comprising a biologically active compound. 
     
     
       7. The lipid nanoparticle of  claim 6 , wherein the biologically active compound is selected from the group consisting of: a small molecule, a peptide, a protein, a carbohydrate, a nucleic acid, and a lipid. 
     
     
       8. The lipid nanoparticle of  claim 4 , wherein the cationic lipid is a lipid comprising a quaternary amine with a nitrogen atom having four organic substituents. 
     
     
       9. The lipid nanoparticle of  claim 4 , wherein the helper lipid is a neutral zwitterionic lipid. 
     
     
       10. The lipid nanoparticle of  claim 5 , wherein the hydrophobic small molecule is selected from the group consisting of a sterol and a hydrophobic vitamin. 
     
     
       11. The lipid nanoparticle of  claim 10 , wherein the hydrophobic small molecule is cholesterol. 
     
     
       12. The lipid nanoparticle of  claim 7 , wherein the protein is selected from the group consisting of a nucleoprotein, a mucoprotein, a lipoprotein, a synthetic polypeptide, a small molecule linked to a protein, and a glycoprotein. 
     
     
       13. The lipid nanoparticle of  claim 7 , wherein the nucleic acid is in the form of a single stranded or partially double stranded oligomer or a polymer composed of ribonucleotides. 
     
     
       14. The lipid nanoparticle of  claim 7 , wherein the nucleic acid is selected from the group consisting of miRNA, antisense oligonucleotides, siRNA, immune-stimulatory oligonucleotides, aptamers, ribozymes, and plasmids encoding a specific gene or siRNA. 
     
     
       15. A pharmaceutical composition comprising a cyclic amino-lipid according to  claim 1 . 
     
     
       16. A pharmaceutical composition comprising a lipid nanoparticle according to  claim 3 . 
     
     
       17. The pharmaceutical composition of  claim 16 , further comprising a biologically active compound. 
     
     
       18. The pharmaceutical composition of  claim 17 , wherein the biologically active compound is a nucleic acid. 
     
     
       19. The pharmaceutical composition of  claim 18 , wherein the nucleic acid is in the form of a single stranded or partially double stranded oligomer or a polymer composed of ribonucleotides. 
     
     
       20. A method of treating a disease that is caused by the over-expression of one or several proteins in a subject, said method comprising the administration of a lipid nanoparticle according to  claim 6  to the subject, wherein the biologically active compound is a nucleic acid, and wherein treatment refers to alleviation of symptoms, decreasing the rate of disease progression, amelioration or palliation of the disease state, remission or improved prognosis, or slowing the progression of the disease. 
     
     
       21. A method of treating a disease that is caused by a reduced, suppressed or missing expression of one or several proteins in a subject, said method comprising administration of a lipid nanoparticle according to  claim 6  to the subject, wherein the biologically active compound is a nucleic acid, and wherein treatment refers to alleviation of symptoms, decreasing the rate of disease progression, amelioration or palliation of the disease state, remission or improved prognosis, or slowing the progression of the disease. 
     
     
       22. A method for generating an immune response in a subject, said method comprising the administration of the lipid nanoparticle according to  claim 6  to the subject, wherein the biologically active compound is an immune-stimulatory nucleic acid.